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Behrouz Zandieh Doulabi

Dear Alessio,

We are using for almost 10 years b-actin clone AC-74 from sigma (A2228 SIGMA) for ASC protein lysates 1:2000-5000 dilution and never have problems.

Good Luck

Daniel M Cohen

beta-actin is highly abundant. Maybe you are over-loading your gel, or not diluting your antibody sufficiently. Not sure which SCBT antibody you're using, but I would recommend the C4 monoclonal for blotting.

Dawn Fischer

I had a similar problem with patient tumor lysates. The actin I had been using from Cell Signaling (always with great results) was giving me a hit and miss signal. Some patient lysates just wouldn't show up. I tried an actin ab from Santa Cruz and then I had a full signal for the very same lysates. I'm assuming there were just differential binding. Try another antibody clone.